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adjuvant and antigen vector systems. Vaccine 11 , 1179–1184. Czerkinsky C. Cholera toxin and cholera B subunit as oralmucosal adjuvant and antigen vector systems
. Clone libraries were constructed based on blue–white selection method [ 24 ]. The purified PCR products were ligated into TA-cloning vectors (pGEM-T Vector System, Promega, USA) than transformed into competent Escherichia coli JM109 cells. To retrieve
manufacturer’s recommendations. Cloning of erp, hspR, lppX, mmaA4 , and ompA genes The PCR products were first ligated to pGEM-T Easy Vector System (Promega, WI, USA) according to manufacturer’s recommendations, and
pGem ® -T Vector System (Promega, Madison, USA) according to manufacturer’s instructions. Insert sequences were amplified using M13 primer sets [ 42 ] followed by nested PCR reaction with the applied specific primers (27F-519R, A340F-A934R, amoA –F1
